Animal feed supply

ABSTRACT

AN ANIMAL FEED SUPPLY, WHICH CONTAINS A SMALL AMOUNT OF THE CULTURED PRODUCTS OBTAINED FROM A STRAIN OF MOLD FUNGI BELONGING TO THE GENUS FUSARIUM AND SELECTED FROM THE GROUP CONSISTING OF FUSARIUM SPLENDENS, FUSARIUM MONILIFORME, FUSARIUM OXYSPORUM AND FUSARIUM SOLANI.

United States Patent O 3,749,580 ANllVIAL FEED SUPPLY Jinnosuke Abe,Kango Miyauchi, and Masatoshi Kanoh, Ohhito-machi, Japan, assignors toToyo Jozo Kabushiki Kaisha, Takata-gun, Shizuoka-ken, Japan No Drawing.Continuation-impart of abandoned application Ser. No. 714,019, Mar. 18,1968. This application Dec. 1, 1970, Ser. No. 94,183

Claims priority, application Japan, Mar. 30, 1967, 42/ 19,517 Int. Cl.A231: 1/00 U.S. Cl. 99-9 4 Claims ABSTRACT OF THE DISCLOSURE An animalfeed supply, which contains a small amount of the cultured productsobtained from a strain of mold fungi belonging to the genus Fusarium andselected from the group consisting of Fusarium splendens, Fusariummoniliforme, Fusarium oxysporum and Fusarium solam'.

CROSS-REFERENCE TO RELATED APPLICATION This is a continuation-in-part ofUS. application Ser. No. 714,019, filed Mar. 18, 1968, by the sameinventors, and now abandoned.

BACKGROUND OF THE INVENTION Field of the invention This inventionrelates to an improvement in feed supplies for domestic animals, birdsand the like.

Description of the prior art Domestic and cultured animals and birdsnaturally constitute an important protein source for human beings.

There are a large number of research treatises on strengthened feedsupplied which have added thereto only vitamins, minerals and/orantibiotic substances. Other proposals have been directed tostrengthening a low grade feed supply by adding a digesting enzyme orthe like thereto. According to another proposal, a low grade feed supplyis subjected to specific treatments to enrich the feed.

The main object of the present invention is to provide an improved feedsupply which is capable of considerably accelerating the growing rate ofdomestic animals such as Oxen, cows, horses, pigs, rabbits, sheep, goatsand the like, in comparison to regular feed fed to animals.

There is one prior suggestion to use culture products of mold fungibelonging to the genus Fusarium as an animal feed, according to whichGibberellin is used as the additive (US. Pat. No. 2,943,938, De Zeeuw etal.). According to a further propasal, estrogenic compounds are admixedwith the feed (US. Pat. No. 3,196,019, Andrews et al.). It has beenfurther proposed to use the mycelial cake of Gibberella fujikuroi forsimilar purposes (U.S. Pat. No. 2,913,340, Chornock et al.).

It should be noted that the strains of the genus Fusarium used in themanufacture of animal feed according to the present invention do notproduce any Gibberellin and/or estrogenic compounds, which is aconsiderable difference from conventional feeds utilized in the priorart. A further difference between the present invention and the priorart is that the animal feed in the former does not contain mycelialcake.

SUMMARY OF THE INVENTION The present invention provides an animal feedcomprising a normal feed containing, as a supplement, the cultureproducts of a strain of mold fungi belonging to the genus Fusarium andselected from the group consisting of Fusarz'um splendens, Fusariummoniliforme, Fusarium oxysporum and Fusarium solani. The quantity of theculture products added to the feed depends naturally upon the kind andnature of animals to be fed, and also upon the specific and occasionalculture stage of the living stock. Generally speaking, however, theamount of culture products added to the feed will range from 0.05 to 5weight percent, preferably from 0.1 to 1.0 weight percent, based uponthe dry weight of the raw feed, in order to obtain the desired results.

DESCRIPTION OF THE PREFERRED EMBODIMENTS In the manufacture ofthe'strengthened feed supply of the present invention, mold fungibelonging to the genus Fusarium are cultured in a liquid culture medium.

In the industrial manufacture of the strengthened feed supply accordingto this invention, the following process may preferably be adopted.

The mold fungi are cultured in a liquid culture medium, the fungusmycelia are removed therefrom, the filtrate is, after adding the propersupplementary additives such as malt dextrin and, the like, spray-driedto provide a powder product which is ready for use.

The culture product as obtained in the aforementioned manner by use ofmold fungi belonging to the genus Fusarium is then added to the feedmaterial, so as to produce the strengthened feed supply proposed by thepresent invention. Generally speaking, the powder-like product referredto above, obtained by spray-drying the culture medium filtrate, afterbeing admixed with a proper quantity of malt dextrin, may be added as isto the raw feed material, when the latter is a solid mixture, in aproper ratio as will become more clear hereinafter. If the feed is in aliquid state, such as, for instance, arti- Eficial milk, the powder likeproduct is either dissolved or suspended in the liquid material.

The structure of the effective substance contained in the aforementionedculture products contained in the filtrate from the cultured liquidmedium and removed of the fungus bodies has not yet been determined. Itis however believed that the substance is a kind of nondialyticprotein-like high molecular weight substance containing effectiveamounts of carbon, hydrogen and nitrogen sources, as well as othergrowth factors for the fungi. The isoelectric point of the substance isin the acidic pH range. The substance can be precipitated from thenutrient liquid medium by the addition of a water-miscible organicsolvent such as alcohol, acetone or the like. This effective substanceis soluble in water, but insoluble in various organic solvents. When thestrengthened feed according to this invention is fed to domesticanimals, the growth rate thereof will increase from 20 to 50% incomparison with the case where a non-strengthened regular feed is fed tothe animals.

As an example, the female pig will generally deliver eight piglets aftera pregnancy of about days. The natural nursing period after delivery isabout 60 days. Since it is possible to give birth twice per year,however, a 60 day nursing period imposes a considerably heavy load uponthe female which may cause physical asthemia and invite adverse effectson the next succeeding pregnancy. Therefore, the piglets must be fedwith artificially regenerated milk, generally after a lapse of time of30-40 days after delivery. The feed is then changed from regeneratedmilk to a certain weaning feed for piglets at about 60-70 days which isthence gradually converted into the normal adult feed supply. Frequentscours will be encountered in these feed-change periods and the growthof the piglets may substantially stop or may be otherwise adverselyaffected.

EXAMPLE 1 1) Preparation of culture products of mold fungi, F usariummom'liforme Mold fungi, Fusarium moniliforme, were inoculated in a 100ml. Waksman culture medium, sterilized and placed in a 500 ml. conicalflask and shake-cultured at 30 C. for 24 hours. The cultured medium wasthen transferred into a liter jar fermenter, containing a sterilizedliquid culture medium (pH being 7.0, measured in advance ofsterilization), said medium comprising: 5% glucose, 2% corn steepliquor, 0.2% calcium carbonate and 0.05% potassium dihydrogen phosphateand aeratingly fermented at 30 C. for 30 hours. The broth wastransferred into a 500 liter fermentation tank, sterilized at 120 C. forminutes, containing 250 liters of a liquid fermentation mediumcomprising: 2% glucose,

final fermentation, fungus mycelia were filtered off and the filtratewas concentrated in vacuo'from 145m. to 1 m.

200 kg. of malt dextrin were added to the concentrated product, agitatedthoroughly and finally spraydried to provide 380 kg. of culturedproducts.

(2) First example of breeding piglets To regenerated milk, prescribed inJ.A.S.Standards B, 0.5% of the above fermentation product from Fusariummoniliforme was added, and the strengthened feed milk was fed to six35-day-old piglets, of Yorkshire origin, three male and three female,for days. These piglets were fed for an additional 33 days with acommerc'ially-available mixed feed for this purpose, containing 34%wheat bran, 22% corn, 7.3% soya bean cake, 7% fish meal, 1.5% calciumcarbonate, 0.5%

secondary calcium phosphate, 0.5% sodium chloride,

0.1% minerals, 0.05% vitamins and 0.05% antibiotics with 15.3% DCP(direct crude protein) and 68.2% TDN (total direct nitrogen), havingadded thereto 0.5% of the fermentation product as obtained above, theoverall feeding period thus amounting to 9 weeks.

In comparison with the above, 3 male and 3 female piglets weresimultaneously fed with a similar feed, yet having none of thefermentation product of the present invention.

The results are shown in the following Tables 1 and 2.

TABLE 1.CONSUMED FEED QUANTITY AND FEED DEMAND RATIO Initial Last WeightConsumed No. of weight weight Weight increase Growth feed Feed pigletsbefore after increase per day ratio Growth quantity demand tested test(kg) test (kg) (kg) (percent) (percent) index kg.) ratio N ormally-fedpiglets 6 6. 58 19. 90 13. 32 211 202. 4 100 25. 433 2. 66 Piglets fedWith strengthened feed 6 6. 24 30.70 24. 46 388 391. 9 193 00.041 2.05

No'rns:

Weight increase Growth ratio= 100.

Initial weight before test Consumed feed quantity Feed demand ratio=-.Weight increase TABLE 2 Period Initiation After After After After AfterAfter After After After of feeding 7 days 14 days 21 days 28 days days42 days 49 days 56 days 63 days Normally fed piglets:

Weight (kg) 6. 58 6. 6. 75 8.18 9. 37 11. 13. 53 15. 53 17. 38 19. 90:!:0. 64 :l:0. 34 10. Ga 0. 69 i0. 63 :l:0. 78 :|:3. 25 :l:3. 88 i5. 245:6. 90 Growth rat-1o (percent)- 5. 31 19. 14 15. 63 18. 38 15. 01 14 2511. 24 13. 51 Piglegs ted with strengthened fee W eight (kg) 6. 24 6. 668. 24 10. 34 12. 52 15. 40 18. 90 22. 31 26. 08 30. :|:0. 67 i0. 76:|:1. 03 1. 17 :l:1. 22 :l:1. 21 5:3. 72 :l:4. 22 5:4. 36 :l:5. 43Growth ratio (percent) 6. 51 21. 20 22. 60 19. 07 20. 63 20. 46 1G. 5415. 58 16. 2

NOTES;

Growth ratio 1+ 2) W applies to the piglet at the time of weighttesting; W1 specifies the piglets Weight at the next prior weighttesting.

(3) Second breeding example of piglets To the aforementioned regeneratedmilk was added 0.5% and 1%, respectively, of the culture productobtained from Fusarium monz'liforme used in the foregoing Example 1-(1),and this intensified feed was fed to 6 piglets, 3 being male and 3 beingfemale, of Berkshire 5 origin for 35 days after 50 days from theirdelivery. The

results are shown in the following Tables 3 and 4.

TABLE 3 Weight-increase (average weight shown by kgs.)

Initiation After After Alter After After of feeding 7th day 14th day21st day 28th day 35th day Fed with feed containing:

1% cultured products 6.55 6. 8. 00 11. 20 12. 5 13. 0 0.5% culturedproducts. 7.60 8.70 9. 60 12.0 15.8 16.3 Normally fed 7. 10 7. 30 8. 15i). 60 10. 3 ll. 7

TABLE 4.CONSUMED FEED QUANTITY AND FEED DEMANDED RATIO Consumed Numberof Initial Last wt. Wieght Growth Growth feed Feed piglets wt. beforeafter test increase ratio/ index quantity demand under test test (kgs.)(kgs.) (kgs.) day (g.) (percent) (kgs.) ratlo Feed containing:

1% cultured products 2 6. 55 12.0 6. 45 184 98.4 10. 35 1. 60 0.5%cultured products..- 2 7. (i 16. 3 8. 7 249 114. 4 11. 45 1. 32 Normalfeed 2 7. 11.7 4. 60 131 64.8 10.60 2.

(4) Third example of feeding piglets addition of said fermentationproduct thereto was fed to a The culture product obtained in theforegoing Example l-(1) from Fusarium monilz'forme was added to acommercially available piglet-breeding feed in an amount of 0.1%, thefeed having a composition of 30.3% wheat bran; 15.0% defatted rice bran;23.0% corn starch; 15.0% barley; 7.0% soya bean cake; 7.0% fish meal;1.5% calcium carbonate; 0.5% secondary calcium phosphate; 0.5 sodiumchloride; 0.1% minerals; 0.05% vitamins and 0.05 antibiotics; 14.45%D.C.P. and 67.38%

second group of six piglets.

In these test feedings, two groups of the same number of piglets derivedfrom a common father pig and two different mother pigs, were fed inorder to obviate hereditary and unexpectable influences. The testresultsare shown in the following Table 6. As seen in the table, thepiglets fed with the strengthened feed supply showed a 18-25% meanshigher growth increase than the other piglets when measured after 60days from the initiation of the feed test.

TABLE 6 After- Initiation of feeding 7 days 14 days 21 days 28 days days42 days 49 days 56 days 60 days Normally fed:

Wt. (kg) 14. 85 17. 68 20. 05 21. 33 22. 85 25. 03 27. 75 30. 95 35. 37.78 84 i=3. 16 i2. 91 =|=1. 81 5:2. 64 11:4. 79 i5. 91 5:6. 32 i6. 70=|;6. 49 Growth ratio (percent)- 17. 37 12. 59 6. 16 6. 91 9. 08 10. 3210. 90 6. 49 0.5% Strengthendly fed:

Wt. (kg) 16. 06 20. 22. 47 26. 13 28. 43 42. 26 36. 23 39. 20 43.0745.03 5:2. 28 i5. i4. 79 i6. 78 i7. 21 i8. 18 i8. 94 i8. 60 i8. 83 i9.43 Growth ratio (percent). 24 24 9 15 15.08 .8. 43 7 86 4. 46

NOTES:

Wr-W Growth ratio 100 W2 indicates the piglet at the time of weighttesting; W indicates the weight of the prior weight testing.

T.D.N. 4 piglets of Berkshire origin were fed with the strengthenedfeed. The results are shown in Table 5.

TABLE 5 Item Wt. Consumed Number of Initial Wt. Last wt. Wt. increaseGrowth feed Feed piglets before test after test increase per day ratioGrowth quantity demand Group under test (kgs.) (kgs.) (kgs.) (g.)(percent) index (kgs.) ratio Normally fed 4 14. 37. 78 22. 93 382 154. 4100 90. 6 3. 95 Fed with strengthened feed 4 16.06 45. O3 28. 97 483180. 3 117 83. 7 2. 89

EXAMPLE 2 EXAMPLE 3 ml. of 1%-glucose buillon was charged in a 500 ml.50

conical flask, sterilized at 120 C. for 20 minutes, and inoculated withFusarium moniliforme, and then shakecultured at 30 C. for 24 hours. Thiscultured medium was transferred to a sterilized jar fermenter,containing 20 liters of a liquid nutritious medium comprising 3.5%glucose; 1.5% corn steep liquor; 0.3% ammonium sulfate; 0.2% calciumcarbonate; 0.05 potassium dihydrogen phosphate and 0.05 magnesiumsulfate, and fermented under aeration at 30 C. for 30 hours. Thethuscultured medium was further transferred to a 500 liter fermentationtank containing 200 liters of a liquid nutritious medium comprising 5%wheat bran; 0.2% ammonium sulfate and 0.1% calcium carbonate andfermented at 30 C. for 24 hours. The cultured broth was further mixedwith 10 m? of another liquid nutritious medium comprising 3% rice bran;2% wheat bran; 0.2% ammonium sulfate and 0.1% calcium carbonate in afermentation tank having a capacity of 15 m. and then fermented at 30 C.for 96 hours under aeration. The fungus mycelia was filtered off and 8.6m. of the filtrate was then mixed with kgs. of malt dextrin andspray-dried. 253 kgs. of a powder-like product was obtained.

0.5 of the fermentation product was then added to the same piglets feedsupply as used in Example 1-(2), and six piglets were fed with thisstrengthened feed for 85 days. In comparison, the same feed supplywithout the In place of F usariurm moniliforme, as used in Example1-(1), other mold fungi listed in the following Table 7 were employedseparately, and similar culture products as referred to hereinbeforewere obtained respectively.

TABLE 7 Kind of mold fungi: Yield of cultured product F usariumsplendens 1 330 Fusarium oxysporum 250 Fusarium solani 2 260 1 Maltdextrin added in advance of spray-drying in an amount of 180 kg.

2 Malt dextrin added in advance of spray-drying in an amount of kg.

When the cultured products obtained from the abovespecified mold fungiwere employed in place of that from Fwsarium monilz'forme, and fedsimilarly as in the case of Example 1-(2), a growth increase ratio of20-40%, relative to the comparative reference, was observed.

EXAMPLE 4 The cultured product obtained in the Example 1-(1) was addedto a commercially available chick-breeding feed in an amount of 0.5%,the feed having a composition shown in the following Table 9, and 50chicks, Wellbrine origin, were breed with the above-intensified feed for30 days after 3 days from delivery. Reference chicks were also fed, butwith the comparative non-intensified feed.

All the chicks in the test were fed naturally with a properly selectedamount of green vegetables.

The results are shown in Table 8. As is seen, the growth increase ratioamounted to about 30% relative to the reference.

Composition of feed Corn, wheat, bean cake, heat-treated soy bean cake,gingelly oil cake, fish meal, dried blood powder, wheat bran, defattedrice bran, lusam meal, yeast, calcium, phosphate, calcium carbonate,manganese sulfate, zinc sulfate, sodium chloride, vitamins, U.G.F.,antibiotics, antinematodes and minerals.

7 EXAMPLE 5 The cultured product obtained in Example 1 was added to afeed having the following composition in an amount of 1% and 20 micewere fed therewith. A higher growth increase rate of 15-25% was observedthan in the case of the reference.

Composition of feed (total: 100 g.)

Crude protein g 24.0 Calcium ..-percent 1.0 Crude fat g 3.5 Phosphorpercent 1.0 Magnesium do 0.27 Crude cellulose g 4.5 Grade ash g 6.0Sodium percent 0.3 l Soluble nitrogen g 56.0 Potassium percent 0.85Calcium pantothenate mg 30 Manganese p.p.m 60.00 Niacin mg 80 Iron p.p.m100.00 Folic acid mg 0.2 Calcium phosphor 1.00 Vitamin A I.UCalcium/magnesium 3.00 Vitamin D3 I.U 2 Potassium/ manganese 2.74Vitamin E mg Vitamin B1 mg 7 Vitamin B2 mg 10.0 Vitamin B6 mg.. 4.0Vitamin B12 mg 0.02

EXAMPLE 6 Rats were used instead of the mice in Example 5 and the testprocedures were substantially the same as in Example 2. Similar resultswere obtained as those in Example 5.

EXAMPLE 7 Vitamins A, D, thiamine, r-iboflavine, biotin; pantothenicacid, niacin, folic acid, vitamins B, E, -K, pyridoxine.

Unknown growth factors.

3 Included trace minerals such as Fe, Cu, Co, K, and the like.

Composition of feed (total: g.)

Crude protein g Calcium percent 1.2 Crude fat g 3.0 Phosphor percent0.58 Crude cellulose g 11.5 Magnesium percent 0.22 Crude ash g 8.0Sodium percent 0.32 Soluble nitrogen g 51.0 Potassium percent 1.02Calcium pantothenate mg 20 Manganese p.p.m. 56.25 Niacin mg 70 Ironp.p.m 155.00 Folic acid mg 0.2 Calcium/ phosphor 2.06 Vitamin A I.U 50

Calcium/magnesium 5.45 Vitamin D3 I.U 3600 Potassium/manganese 3. 18Vitamin E mg 15 Vitamin B1 mg 5 Vitamin B2 mg 10 Vitamin B6 rng 4.0Vitamin B12 mg 002 Vitamin C mg' 2.00

The results were such that a growth rate of the rabbits fed with thestrengthened feed was 50% higher than the comparative conventionalfeeding.

EXAMPLE 8 10 rabbits were fed with normal green weeds and I vegetables,of which, however, 5 rabbits were fed for two months with the green feedcontaining the cultured product obtained in Example 2 and dissolved in aphysiological saline solution, said product being dosed at the rate of100 mg./kg./day, or more specifically 5 mL/animal/day. It was found thata 15-20% higher growth ratio was attained than in the reference rabbits.

EXAMPLE 9 1000 ml. of 1%-glucose bullion was equally divided and chargedinto 10 separate conical flasks, each having a 500 ml. capacity, andupon regular sterilization, inoculated with strains of Fusariummoniliferme, and then shakecultured at 30 C. for 24 hours. Theinoculating liquor thus prepared was introduced into a fermentationtang, of 200 liter capacity, containing, after being sterilized at C.for 20 minutes, 100 liters of a nutritious liquid medium, and thenfermented at 30 C. for 96 hours under aeration. The thus cultured brothwas filtered and alcohol in an amount of 5 times the amount present wasadded under agitation. The resulting precipitate dried to 500 g. of apowder-like product as before.

This powder was added to a conventional commercial feed supply forpiglets of Berkshire origin in an amount of 0.5% for 60 days after 60days from the delivery, the feed supply being of the same compositiondisclosed in Example 2. The test results were such that the piglets .fedwith the strengthened feed showed a 23%-higher growth ratio in the meanthan the reference piglets fed with the comparative regular feed.

EXAMPLE 10 Two calves of Holstein origin were fed with a regular feed,strengthened with 0.5% of the cultured product obtained in Example 1 for15 months. The thus fed calves had 375 kg. of mean grown weight, whilethe comparative reference animals showed only 330 kg. of mean weightgain.

The present invention may be embodied in other specific forms withoutdeparting from the spirit or essential characteristics thereof. Thepresent embodiments are therefore to be considered in all respects asillustrative and not restrictive, the scope of the invention beingindicated by the appended claims rather than by the foregoingdescription, and all changes which come within the meaning and range ofequivalency of the claims are therefore intended to be embraced therein.What is claimed is: 1. An animal feed composition, comprising, inadmixture:

(1) a conventional animal feed, and (2) an effective quantity of a driedproduct free of fungus mycelia, obtained from the aerobic cultivation,in a liquid nutrient medium, sufiicient to sustain normal growth, of agibberellin non-producing strain of mold fungi belonging to the genusFusarium and selected from the group consisting of F usarium splendens,F usarium oxysporum and Fusarium solani. 2. The animal feed compositionof claim 1 wherein the dried product is added to the feed in an amountof from 0.1 to 1% by weight, based on the dry Weight of the conventionalfeed.

3. The animal feed composition as in claim 1 wherein the dried productis added to the feed in an amount of from 0.05% to 5.0% by weight, basedon the dry weight of the conventional feed.

4. The animal feed composition as in claim 3 wherein the dried productis in the form of a white powder spraydried and obtainable from theliquid medium of the cultivation of said mold fungi.

References Cited UNITED STATES PATENTS 2,913,340 11/1959 Chornock et a1992 2,450,055 9/1948 Nord 195- -8l 3,085,049 4/1963 Rudy et a1. 19'5-80A. LOUIS MONACELL, Primary Examiner R. M. ELLIOTT, Assistant ExaminerU.S. C1. X.R. 992

